Western blot using affinity purified anti-C-Myb antibody in Cos7 cell lysates: Lane 1: vector; Lane 2: transfected with c-myb, no treatment; Lane 3: transfected with c-Myb and heat stress-treated (incubation at 42°C for 30minutes) to induce c-Myb modification. Lane 3 shows detection of bands at 75, 98, and 125kDa corresponding to overexpressed c-Myb (arrows): wild-type, with one attached SUMO molecule, and with two attached SUMO molecules, respectively. Nitrocellulose 0.45µm membrane was blocked for 1 hr at RT in 10% BLOTTO in PBS. The primary antibody was used at 1:500.
